Serrano Pepper Hybrid XHP16823

ABSTRACT

One embodiment relates to seed and plants of pepper hybrid XHP16823. Another embodiment relates to the plants, seeds and tissue cultures of pepper hybrid XHP16823, and to methods for producing a pepper plant produced by crossing such plants with themselves, with another pepper plant, such as a plant of another genotype, or with vegetatively propagating said plant. Another embodiment further relates to seeds and plants produced by such crossing. Further embodiments relate to parts of such plants, including the fruit and gametes of such plants.

BACKGROUND

The embodiments recited herein relates to a novel and distinct serranohybrid pepper (Capsicum annuum) designated XHP16823, and to the seeds,plant parts, and tissue culture produced by that hybrid pepper. Theembodiments further relate to food products produced from hybrid pepperXHP16823, such as, but not limited to, fruit, powders, sauces, andsalsas. All publications cited in this application are hereinincorporated by reference.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

BRIEF DESCRIPTION OF THE FIGURES

The patent or application file may contain one or more drawings executedin color and/or one or more photographs. Copies of this patent or patentapplication publication with color drawing(s) and/or photograph(s) willbe provided by the Patent Office upon request and payment of thenecessary fee.

FIG. 1 shows an individual fruit of serrano hybrid pepper XHP16823.

FIG. 2 shows an individual fruit of the commercial serrano comparison,‘Camino Real’.

SUMMARY

It is to be understood that the embodiments include a variety ofdifferent versions or embodiments, and this Summary is not meant to belimiting or all-inclusive. This Summary provides some generaldescriptions of some of the embodiments, but may also include some morespecific descriptions of other embodiments.

An embodiment provides a hybrid pepper designated XHP16823. Anotherembodiment relates to the seeds of hybrid pepper XHP16823, to the plantsof hybrid pepper XHP16823 and to methods for producing a pepper plantproduced by crossing hybrid pepper XHP16823 with itself or anotherhybrid pepper, and the creation of variants by mutagenesis ortransformation of hybrid pepper XHP16823.

Any such methods using hybrid pepper XHP16823 are a further embodiment:selfing, backcrosses, hybrid production, crosses to populations, and thelike. All plants produced using hybrid pepper XHP16823 as at least oneparent are within the scope of the embodiments. Advantageously, hybridpepper XHP16823 could be used in crosses with other, different pepperplants to produce first generation (F₁) pepper hybrid seeds and plantswith superior characteristics.

Another embodiment provided for a plant of hybrid pepper comprising anadded heritable trait is provided. The heritable trait may comprise agenetic locus that is, for example, a dominant or recessive allele. Inanother embodiment, a plant of hybrid pepper XHP16823 is defined ascomprising a single locus conversion. In another embodiment, an addedgenetic locus confers one or more traits such as, for example, herbicidetolerance, insect tolerance, tolerance for bacterial, fungal, or viraldisease, male fertility, male sterility, enhanced nutritional quality,modified carbohydrate metabolism, environmental stress tolerance,modified yield, and industrial usage. In further embodiments, the traitmay be conferred by a naturally occurring gene introduced into thegenome of a line by backcrossing, a natural or induced mutation, or atransgene introduced through genetic transformation techniques into theplant or a progenitor of any previous generation thereof. Whenintroduced through transformation, a genetic locus may comprise one ormore genes integrated at a single chromosomal location.

Another embodiment provides for regenerable cells for use in tissueculture of hybrid pepper XHP16823. The tissue culture may be capable ofregenerating plants having all the physiological and morphologicalcharacteristics of the foregoing pepper plant, and of regeneratingplants having substantially the same genotype as the foregoing pepperplant. The regenerable cells in such tissue cultures may be embryos,protoplasts, meristematic cells, callus, pollen, leaves, ovules,anthers, cotyledons, hypocotyl, pistils, roots, root tips, fruit,flowers, seeds, plant, petiole, or stems. Still a further embodimentprovides for pepper plants regenerated from the tissue cultures ofhybrid pepper XHP16823.

Another embodiment relates to a method of vegetatively propagatingpepper plant XHP16823 comprising the steps of: (a) collecting tissuecapable of being propagated from a XHP16823 plant; (b) cultivating saidtissue to obtain proliferated shoots; and (c) rooting said proliferatedshoots to obtain rooted plantlets.

Another embodiment provides for the pepper seeds and plants produced bya process that comprises crossing a first parent pepper plant with asecond parent pepper plant, wherein at least one of the first or secondparent pepper plants is a plant of hybrid pepper hybrid. In oneembodiment of the invention, pepper seed and plants produced by theprocess are first generation (F₁) hybrid pepper seed and plants producedby crossing a plant in accordance with the embodiments, with another,distinct plant. Another embodiment further contemplates plant parts ofsuch an F₁ hybrid pepper plant, and methods of use thereof. Therefore,further embodiments provide for an F₁ hybrid pepper plant and seedthereof. Plant parts (or a pepper plant, or a part thereof) includes butis not limited to, regenerable cells in such tissue cultures may beembryos, protoplasts, meristematic cells, callus, pollen, anther, shoot,leaves, ovules, anthers, cotyledons, hypocotyl, pistils, roots, roottips, fruit, flowers, seeds, plant, petiole, or stems. A cotyledon is atype of seed leaf. The cotyledon contains the food storage tissues ofthe seed. The embryo is the small plant contained within a mature seed.A hypocotyl is the portion of an embryo or seedling between thecotyledons and the root. Therefore, it can be considered a transitionzone between shoot and root.

Another embodiment provides for a method of producing a plant derivedfrom hybrid pepper XHP16823, the method comprising the steps of: (a)preparing a progeny plant derived from hybrid pepper XHP16823, whereinsaid preparing comprises crossing a plant of hybrid XHP16823 with asecond plant; and (b) crossing the progeny plant with itself or a secondplant to produce a seed of a progeny plant of a subsequent generation.In further embodiments, the method may additionally comprise: (c)growing a progeny plant of a subsequent generation from said seed of aprogeny plant of a subsequent generation and crossing the progeny plantof a subsequent generation with itself or a second plant; and repeatingthe steps for an additional 3-10 generations to produce a plant derivedfrom hybrid pepper XHP16823. The plant derived from hybrid XHP16823 maybe an inbred line, and the aforementioned repeated crossing steps may bedefined as comprising sufficient inbreeding to produce the inbred line.In the method, it may be desirable to select particular plants resultingfrom step (c) for continued crossing according to steps (b) and (c). Byselecting plants having one or more desirable traits, a plant derivedfrom hybrid XHP16823 is obtained which possesses some of the desirabletraits of the line/hybrid as well as potentially other selected traits.

Further embodiments provide for a method of producing food or feedcomprising: (a) obtaining a plant of hybrid pepper XHP16823, wherein theplant has been cultivated to maturity, and (b) collecting at least onepepper from the plant.

In another embodiment, the first step in “crossing” comprises plantingseeds of a first and second parent pepper plant, often in proximity sothat pollination will occur for example, mediated by insect vectors.Alternatively, pollen can be transferred manually. Where the plant isself-pollinated, pollination may occur without the need for direct humanintervention other than plant cultivation. A second step may comprisecultivating or growing the seeds of first and second parent pepperplants into plants that bear flowers. A third step may comprisepreventing self-pollination of the plants, such as by emasculating theflowers (i.e., killing or removing the pollen). A fourth step for ahybrid cross may comprise cross-pollination between the first and secondparent pepper plants. Yet another step comprises harvesting the seedsfrom at least one of the parent pepper plants. The harvested seed can begrown to produce a pepper plant or hybrid pepper plant.

Another embodiment provides for a method for developing a pepper plantin a plant breeding program, comprising applying plant breedingtechniques to the plant, or plant part thereof, of pepper XHP16823,comprising crossing, recurrent selection, mutation breeding, whereinsaid mutation breeding selects for a mutation that is spontaneous orartificially induced, backcrossing, pedigree breeding, marker enhancedselection, haploid/double haploid production, or transformation topepper plant XHP16823, or its parts, wherein application of saidtechniques results in development of a pepper plant.

Another embodiment provides for a method of introducing a mutation intothe genome of pepper plant XHP16823, said method comprising mutagenesisof the plant, or plant part thereof, of pepper plant XHP16823, whereinsaid mutagenesis is selected from the group consisting of temperature,long-term seed storage, tissue culture conditions, ionizing radiation,chemical mutagens, or targeting induced local lesions in genomes, andwherein the resulting plant comprises at least one genome mutation.

Another embodiment provides for a method of editing the genome of pepperplant XHP16823, said method comprising editing the genome of the plant,or plant part thereof, of pepper plant XHP16823, wherein said method isselected from the group comprising zinc finger nucleases, transcriptionactivator-like effector nucleases (TALENs), engineered homingendonucleases/meganucleases, and the clustered regularly interspacedshort palindromic repeat (CRISPR)-associated protein9 (Cas9) system.

The pepper seed of XHP16823 may be provided as an essentiallyhomogeneous population of pepper seed of pepper hybrid XHP16823.Essentially homogeneous populations of seed are generally free fromsubstantial numbers of other seed.

Another embodiment provides for the genetic complement of hybrid pepperXHP16823. The phrase “genetic complement” is used to refer to theaggregate of nucleotide sequences, the expression of which sequencesdefines the phenotype of, in the present case, a pepper plant, or a cellor tissue of that plant. A genetic complement thus represents thegenetic makeup of a cell, tissue or plant, and a hybrid geneticcomplement represents the genetic make-up of a hybrid cell, tissue orplant. Thus, another embodiment provides for pepper plant cells thathave a genetic complement in accordance with the pepper plant cellsdisclosed herein, and seeds and plants containing such cells.

As used herein, “at least one,” “one or more,” and “and/or” areopen-ended expressions that are both conjunctive and disjunctive inoperation. For example, each of the expressions “at least one of A, Band C,” “at least one of A, B, or C,” “one or more of A, B, and C,” “oneor more of A, B, or C” and “A, B, and/or C” means A alone, B alone, Calone, A and B together, A and C together, B and C together, or A, B andC together.

Various embodiments are set forth in the Detailed Description asprovided herein and as embodied by the claims. It should be understood,however, that this Summary does not contain all of the aspects andembodiments, is not meant to be limiting or restrictive in any manner,and that embodiment(s) as disclosed herein is/are understood by those ofordinary skill in the art to encompass obvious improvements andmodifications thereto.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION

Hybrid pepper XHP16823 is a serrano pepper that is adapted to most areasin the United States. XHP16823 has a compact plant habit and a hotpungency.

Hybrid pepper XHP16823 has shown uniformity and stability, as describedin the following variety description information. Hybrid pepper XHP16823was tested for uniformity and stability a sufficient number ofgenerations with careful attention to uniformity of plant type and hasbeen increased with continued observation for uniformity.

Hybrid pepper XHP16823 has the following morphologic and othercharacteristics based primarily on data collected in Woodland, Calif.RHS refers to the Royal Horticultural Society color reference. Wall orflesh thickness refers to the diameter of the fruit wall. Wall thicknessis measured by cutting the fruit in half longitudinally and measuringthe thickness (mm) on the thinnest part of each side of the wall of eachfruit. Wall thickness is important to both bell pepper growers andshippers because fruit having thick walls are more likely to ship betterwithout the fruit wall cracking.

TABLE 1 TABLE 1: VARIETY DESCRIPTION INFORMATION Characteristic XHP16823General Fruit Type Serrano (small hot) Adaptation Most U.S. Areas PlantHabit Compact Plant Attitude Semi-Erect Plant Height 51.0 cm Plant Width56.0 cm Length of Stem From Cotyledons to 1st Flower 13.0 cm Length ofThird Internode (from soil surface) 90.0 mm Basal Branches Many (morethan 3) Branch Flexibility Rigid Stem Strength Strong Mature Leaf ShapeLanceolate Leaf Length 27.0 mm Leaf width 50.0 mm Petiole Length 40.0 mmLeaf and Stem Pubescence Absent Margin Undulation Weak Blistering AbsentLeaf Color Medium Green, RHS 137A Number of Flowers Per Leaf Axil 3Number of Calyx Lobes 6 Number of Petals 6 Flower Diameter 16.0 mmCorolla Color White Corolla Throat Markings Yellow (Tan) Anther ColorPurple Style Length Exceeds stamen Self-Incompatibility Absent ImmatureFruit Color Light Green Mature Fruit Color Red Pungency Hot FruitGlossiness Shiny Surface Smoothness Smooth Fruit Position Pendant CalyxShape Cup-Shaped (enveloping fruit base) Calyx Diameter 15.0 mm FruitLength 95.0 mm Fruit Diameter at Calyx Attachment 15.0 mm Fruit Diameterat Mid-Point 13.0 mm Flesh Thickness at Mid-Point 3.0 mm Average Numberof Fruit/Plant 100  Average Fruit Weight 18.0 gram Fruit Base ShapeRounded Fruit Apex Shape Pointed Fruit Shape Oblong Fruit Shape(Longitudinal Section) Horned-shaped Fruit Shape (Cross Section)Circular Fruit Set Concentrated Interloculary Grooves Absent Percentageof fruits with two locules 20  Percentage of fruits with three locules70  Percentage of fruits with four locules 15  Percentage of fruits withfive or more locules 0 Pedicel Length 19.0 mm Pedicel Thickness 3.0 mmPedicel Shape Curved Pedicel Cavity Absent Seed Color Yellow AnthocyaninPresence in Seedling Hypocotyl Absent Anthocyanin Presence in StemAbsent Anthocyanin Presence in Node Weak Anthocyanin Presence in LeafAbsent Anthocyanin Presence in Pedicel Absent Anthocyanin Presence inCalyx Absent Anthocyanin Presence in Fruit AbsentComparison of Hybrid Pepper XHP16823 with Commercial Variety

Table 2 below is a comparison of hybrid pepper XHP16823 with thecommercial comparison variety ‘Camino Real’.

TABLE 2 COMPARISON OF XHP16823 WITH ‘CAMINO REAL’ Number of Seed CavitySeed Cavity Placenta Seeds per Variety Length (mm) Diameter (mm) Length(mm) Fruit XHP16823 84.8 18.8 62.7 120 Camino Real 39.5 7.2 35.8 35Breeding with Hybrid Pepper XHP16823

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable variety. This approach hasbeen used extensively for breeding disease-resistant varieties. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for three or more years. The best lines are candidatesfor new commercial varieties; those still deficient in a few traits maybe used as parents to produce new populations for further selection.

These processes, which lead to the final step of marketing anddistribution, is a time-consuming process that requires precise forwardplanning, efficient use of resources, and a minimum of changes indirection.

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardvariety. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of pepper breeding is to develop new and superior peppervarieties and hybrids. The breeder initially selects and crosses two ormore parental lines, followed by repeated selfing and selection,producing many new genetic combinations. The breeder can theoreticallygenerate billions of different genetic combinations via crossing,selection, selfing and mutations.

Using Hybrid Pepper XHP16823 to Develop Other Pepper Varieties

Pepper varieties such as hybrid pepper XHP16823 are typically developedfor fresh consumption. However, pepper varieties such as hybrid pepperXHP16823 also provide a source of breeding material that may be used todevelop new pepper varieties. Plant breeding techniques known in the artand used in a pepper breeding program include, but are not limited to,recurrent selection, bulk selection, mass selection, backcrossing,pedigree breeding, open pollination breeding, restriction fragmentlength polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, transformation, and gene editing.These techniques can be used singularly or in combinations. Thedevelopment of pepper varieties in a breeding program requires, ingeneral, the development and evaluation of homozygous varieties. Thereare many analytical methods available to evaluate a new variety. Theoldest and most traditional method of analysis is the observation ofphenotypic traits, but genotypic analysis may also be used.

Additional Breeding Methods

One embodiment is directed to methods for producing a pepper plant bycrossing a first parent pepper plant with a second parent pepper plant,wherein the first or second pepper plant is the pepper plant from hybridpepper XHP16823. Further, both first and second parent pepper plants maybe from hybrid pepper XHP16823. Any plants produced using hybrid pepperXHP16823 as at least one parent are also within the scope of theembodiments. These methods are well known in the art and some of themore commonly used breeding methods are described herein. Descriptionsof breeding methods can be found in one of several reference books(e.g., Allard, Principles of Plant Breeding (1960); Simmonds, Principlesof Crop Improvement (1979); Sneep, et al. (1979); Cooper, S. G., D. S.Douches and E. J. Grafius. (2004).

The following describes breeding methods that may be used with hybridpepper XHP16823 in the development of further pepper plants. One suchembodiment is a method for developing a hybrid pepper XHP16823 progenyplant in a pepper breeding program comprising: obtaining the pepperplant, or a part thereof, of hybrid pepper XHP16823, utilizing saidplant, or plant part, as a source of breeding material, and selecting ahybrid pepper XHP16823 progeny plant with molecular markers in commonwith hybrid pepper XHP16823 and/or with morphological and/orphysiological characteristics selected from the characteristics listedin Table 1. Breeding steps that may be used in the pepper plant breedingprogram include pedigree breeding, backcrossing, mutation breeding, andrecurrent selection. In conjunction with these steps, techniques such asRFLP-enhanced selection, genetic marker enhanced selection (for example,SSR markers), and the making of double haploids may be utilized.

Another method involves producing a population of hybrid pepper XHP16823progeny pepper plants, comprising crossing hybrid pepper XHP16823 withanother pepper plant, thereby producing a population of pepper plantswhich derive 50% of their alleles from hybrid pepper XHP16823. A plantof this population may be selected and repeatedly selfed or sibbed witha hybrid pepper line resulting from these successive filial generations.One embodiment is the hybrid pepper produced by this method and that hasobtained at least 50% of its alleles from hybrid pepper XHP16823.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see, Fehr and Walt, Principles of VarietyDevelopment, pp. 261-286 (1987). Thus, embodiments include hybrid pepperXHP16823 progeny pepper plants comprising a combination of at least twohybrid pepper XHP16823 traits selected from the group consisting ofthose listed in Table 1 and a combination of traits listed in theSummary, so that said progeny pepper plant is not significantlydifferent for said traits than hybrid pepper XHP16823 as determined atthe 5% significance level when grown in the same environmentalconditions. Using techniques described herein, molecular markers may beused to identify said progeny plant as a hybrid pepper XHP16823 progenyplant. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions. Once such avariety is developed, its value is substantial since it is important toadvance the germplasm base as a whole in order to maintain or improvetraits such as yield, disease tolerance, pest tolerance, and plantperformance in extreme environmental conditions.

Progeny of hybrid pepper XHP16823 may also be characterized throughtheir filial relationship with hybrid pepper XHP16823, as for example,being within a certain number of breeding crosses of hybrid pepperXHP16823. A breeding cross is a cross made to introduce new geneticsinto the progeny, and is distinguished from a self or a sib cross, whichis made to select among existing genetic alleles. The lower the numberof breeding crosses in the pedigree, the closer the relationship betweenhybrid pepper XHP16823 and its progeny. For example, progeny produced bythe methods described herein may be within 1, 2, 3, 4, or 5 breedingcrosses of hybrid pepper XHP16823.

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such ashybrid pepper XHP16823 and another hybrid pepper having one or moredesirable characteristics that is lacking or which complements hybridpepper XHP16823. If the two original parents do not provide all thedesired characteristics, other sources can be included in the breedingpopulation. In the pedigree method, superior plants are selfed andselected in successive filial generations. In the succeeding filialgenerations, the heterozygous condition gives way to homogeneousvarieties as a result of self-pollination and selection. Typically, inthe pedigree method of breeding, five or more successive filialgenerations of selfing and selection is practiced: F₁ to F₂; F₂ to F₃;F₃ to F₄; F₄ to F₅; etc. After a sufficient amount of inbreeding,successive filial generations will serve to increase seed of thedeveloped variety. Preferably, the developed variety compriseshomozygous alleles at about 95% or more of its loci.

Backcross Breeding

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous variety orhybrid which is the recurrent parent. The source of the trait to betransferred is called the donor parent. After the initial cross,individuals possessing the phenotype of the donor parent are selectedand repeatedly crossed (backcrossed) to the recurrent parent. Theresulting plant is expected to have the attributes of the recurrentparent (e.g., variety) and the desirable trait transferred from thedonor parent. This is also known as single gene conversion and/orbackcross conversion.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single gene of the recurrent variety ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some agronomically important trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered to determine an appropriate testing protocol.Although backcrossing methods are simplified when the characteristicbeing transferred is a dominant allele, a recessive allele may also betransferred. It may be necessary to introduce a test of the progeny todetermine if the desired characteristic has been successfullytransferred.

A backcross conversion of hybrid pepper XHP16823 occurs when DNAsequences are introduced through backcrossing, with hybrid pepperXHP16823 utilized as the recurrent parent. Both naturally occurring andtransgenic DNA sequences may be introduced through backcrossingtechniques. A backcross conversion may produce a plant with a trait orlocus conversion in at least two or more backcrosses, including at least2 crosses, at least 3 crosses, at least 4 crosses, at least 5 crosses,and the like. Molecular marker assisted breeding or selection may beutilized to reduce the number of backcrosses necessary to achieve thebackcross conversion. For example, see, Frisch M. et al,“Marker-Assisted Backcrossing for Simultaneous Introgression of TwoGenes” Crop Science Society of America, pp 1716-1725 (2001) andOpenshaw, S. J., et al., “Marker-assisted Selection in BackcrossBreeding, Proceedings Symposium of the Analysis of Molecular Data” CropScience Society of America, Corvallis, Oreg. (August 1994), where it wasdemonstrated that a backcross conversion could be made in as few as twobackcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes ascompared to unlinked genes), the level of expression of the trait, thetype of inheritance (cytoplasmic or nuclear), and the types of parentsincluded in the cross. It is understood by those of ordinary skill inthe art that for single gene traits that are relatively easy toclassify, the backcross method is effective and relatively easy tomanage. Desired traits that may be transferred through backcrossconversion include, but are not limited to, sterility (nuclear andcytoplasmic), fertility restoration, nutritional enhancements, droughttolerance, nitrogen utilization, low phytate, industrial enhancements,disease tolerance (bacterial, fungal, or viral), insect tolerance, andherbicide tolerance. In addition, an introgression site itself, such asan FRT site, Lox site, or other site-specific integration site, may beinserted by backcrossing and utilized for direct insertion of one ormore genes of interest into a specific plant variety. In someembodiments, the number of loci that may be backcrossed into hybridpepper XHP16823 is at least 1, 2, 3, 4, or 5, and/or no more than 6, 5,4, 3, or 2. A single locus (locus or loci (plural) refers to a positionin the genome for a gene, SNP, mutation, etc.) may contain severaltransgenes, such as a transgene for disease tolerance that, in the sameexpression vector, also contains a transgene for herbicide tolerance.The gene for herbicide tolerance may be used as a selectable markerand/or as a phenotypic trait. A single locus conversion of site-specificintegration system allows for the integration of multiple genes at theconverted loci.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selfing eachbackcross generation to determine which plants carry the recessivealleles unless molecular markers are available to track the gene.Recessive traits may require additional progeny testing in successivebackcross generations to confirm the presence of the locus of interest.The last backcross generation is usually selfed to give pure breedingprogeny for the gene(s) being transferred, although a backcrossconversion with a stably introgressed trait may also be maintained byfurther backcrossing to the recurrent parent with selection for theconverted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype of the recurrent parent. The backcross is a form ofinbreeding, and the features of the recurrent parent are automaticallyrecovered after successive backcrosses. Poehlman, “Breeding Field Crops”p. 204 (1987). Poehlman suggests from one to four or more backcrosses,but as noted above, the number of backcrosses necessary can be reducedwith the use of molecular markers. Other factors, such as a geneticallysimilar donor parent, may also reduce the number of backcrossesnecessary. As noted by Poehlman, backcrossing is easiest for simplyinherited, dominant, and easily recognized traits.

One process for adding or modifying a trait or locus in hybrid pepperXHP16823 comprises crossing hybrid pepper XHP16823 plants grown fromhybrid pepper XHP16823 seed with plants of another hybrid pepper thatcomprise the desired trait or locus, selecting F₁ progeny plants thatcomprise the desired trait or locus to produce selected F₁ progenyplants, crossing the selected progeny plants with the hybrid pepperXHP16823 plants to produce backcross progeny plants, selecting forbackcross progeny plants that have the desired trait or locus and themorphological characteristics of hybrid pepper XHP16823 to produceselected backcross progeny plants, and backcrossing to hybrid pepperXHP16823 three or more times in succession to produce selected fourth orhigher backcross progeny plants that comprise said trait or locus. Themodified hybrid pepper XHP16823 may be further characterized as havingthe physiological and morphological characteristics of hybrid pepperXHP16823 listed in Table 1 as determined at the 5% significance levelwhen grown in the same environmental conditions and/or may becharacterized by percent similarity or identity to hybrid pepperXHP16823 as determined by SSR markers. The above method may be utilizedwith fewer backcrosses in appropriate situations, such as when the donorparent is highly related or markers are used in the selection step.Desired traits that may be used include those nucleic acids known in theart, some of which are listed herein, that will affect traits throughnucleic acid expression or inhibition. Desired loci include theintrogression of FRT, Lox, and other sites for site specificintegration, which may also affect a desired trait if a functionalnucleic acid is inserted at the integration site.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny pepper seed byadding a step at the end of the process that comprises crossing hybridpepper XHP16823 with the introgressed trait or locus with a differentpepper plant and harvesting the resultant first-generation progenypepper seed.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety but that can beimproved by backcrossing techniques well-known in the art. Single genetraits may or may not be transgenic. Examples of these traits include,but are not limited to, herbicide tolerance, insect tolerance, tolerancefor bacterial, fungal, or viral disease, male fertility, male sterility,enhanced nutritional quality, modified carbohydrate metabolism, modifiedyield, environmental stress tolerance, and industrial usage

In addition to being used to create a backcross conversion, backcrossingcan also be used in combination with pedigree breeding. As discussedpreviously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety, the donor parent, to adeveloped variety called the recurrent parent, which has overall goodagronomic characteristics yet lacks that desirable trait or traits.However, the same procedure can be used to move the progeny toward thegenotype of the recurrent parent, but at the same time retain manycomponents of the nonrecurrent parent by stopping the backcrossing at anearly stage and proceeding with selfing and selection. For example, ahybrid pepper may be crossed with another variety to produce afirst-generation progeny plant. The first-generation progeny plant maythen be backcrossed to one of its parent varieties to create a BC₁ orBC₂. Progeny are selfed and selected so that the newly developed varietyhas many of the attributes of the recurrent parent and yet several ofthe desired attributes of the nonrecurrent parent. This approachleverages the value and strengths of the recurrent parent for use in newpepper varieties.

Therefore, an embodiment of the present disclosure is a method of makinga backcross conversion hybrid pepper XHP16823, comprising the steps ofcrossing a plant of hybrid pepper XHP16823 with a donor plant comprisinga desired trait, selecting an F₁ progeny plant comprising the desiredtrait, and backcrossing the selected F₁ progeny plant to a plant ofhybrid pepper XHP16823 to produce BC₁, BC₂, BC₃, etc. This method mayfurther comprise the step of obtaining a molecular marker profile ofhybrid pepper XHP16823 and using the molecular marker profile to selectfor a progeny plant with the desired trait and the molecular markerprofile of hybrid pepper XHP16823. In one embodiment, the desired traitis a mutant gene, gene, or transgene present in the donor parent.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. Hybrid pepper XHP16823 is suitable foruse in a recurrent selection program. The method entails individualplants cross pollinating with each other to form progeny. The progenyare grown and the superior progeny selected by any number of selectionmethods, which include individual plant, half-sib progeny, full-sibprogeny, and selfed progeny. The selected progeny are cross pollinatedwith each other to form progeny for another population. This populationis planted and again superior plants are selected to cross pollinatewith each other. Recurrent selection is a cyclical process and thereforecan be repeated as many times as desired. The objective of recurrentselection is to improve the traits of a population. The improvedpopulation can then be used as a source of breeding material to obtainnew varieties for commercial or breeding use, including the productionof a synthetic variety. A synthetic variety is the resultant progenyformed by the intercrossing of several selected varieties.

Mass selection is a useful technique when used in conjunction withmolecular marker enhanced selection. In mass selection, seeds fromindividuals are selected based on phenotype or genotype. These selectedseeds are then bulked and used to grow the next generation. Bulkselection requires growing a population of plants in a bulk plot,allowing the plants to self-pollinate, harvesting the seed in bulk, andthen using a sample of the seed harvested in bulk to plant the nextgeneration. Also, instead of self-pollination, directed pollinationcould be used as part of the breeding program.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified, or created,by intercrossing several different parents. The plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Mutation Breeding

Mutation breeding is another method of introducing new traits intohybrid pepper XHP16823. Mutations that occur spontaneously or areartificially induced can be useful sources of variability for a plantbreeder. The goal of artificial mutagenesis is to increase the rate ofmutation for a desired characteristic. Mutation rates can be increasedby many different means including temperature, long-term seed storage,tissue culture conditions, radiation; such as X-rays, Gamma rays (e.g.,cobalt 60 or cesium 137), neutrons, (product of nuclear fission byuranium 235 in an atomic reactor), Beta radiation (emitted fromradioisotopes such as phosphorus 32 or carbon 14), or ultravioletradiation (preferably from 2500 to 2900 nm), or chemical mutagens (suchas base analogues (5-bromo-uracil)), related compounds (8-ethoxycaffeine), antibiotics (streptonigrin), alkylating agents (sulfurmustards, nitrogen mustards, epoxides, ethylenamines, sulfates,sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, oracridines. Once a desired trait is observed through mutagenesis thetrait may then be incorporated into existing germplasm by traditionalbreeding techniques. Details of mutation breeding can be found in Fehr,“Principles of Variety Development,” Macmillan Publishing Company(1993). In addition, mutations created in other pepper plants may beused to produce a backcross conversion of hybrid pepper XHP16823 thatcomprises such mutation.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method, suchas microprojectile-mediated delivery, DNA injection, electroporation,and the like. More preferably, expression vectors are introduced intoplant tissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformant plants obtained with the protoplasm of the embodiments areintended to be within the scope of the embodiments.

Gene Editing Using CRISPR

Targeted gene editing can be done using CRISPR/Cas9 technology (Saunders& Joung, Nature Biotechnology, 32, 347-355, 2014). CRISPR is a type ofgenome editing system that stands for Clustered Regularly InterspacedShort Palindromic Repeats. This system and CRISPR-associated (Cas) genesenable organisms, such as select bacteria and archaea, to respond to andeliminate invading genetic material. Ishino, Y., et al. J. Bacteriol.169, 5429-5433 (1987). These repeats were known as early as the 1980s inE. coli, but Barrangou and colleagues demonstrated that S. thermophiluscan acquire resistance against a bacteriophage by integrating a fragmentof a genome of an infectious virus into its CRISPR locus. Barrangou, R.,et al. Science 315, 1709-1712 (2007). Many plants have already beenmodified using the CRISPR system, including Capsicum annuum. See forexample, U.S. Application Publication No. WO2014068346 (György et al.,Identification of a Xanthomonas euvesicatoria resistance gene frompepper (Capsicum annuum) and method for generating plants withresistance), Martinelli, F. et al., “Proposal of a Genome Editing Systemfor Genetic Resistance to Tomato Spotted Wilt Virus” American Journal ofApplied Sciences 2014, and Noman, A. et al., “CRISPR-Cas9: Tool forQualitative and Quantitative Plant Genome Editing” Frontiers in PlantScience Vol. 7 November 2016.

Gene editing can also be done using crRNA-guided surveillance systemsfor gene editing. Additional information about crRNA-guided surveillancecomplex systems for gene editing can be found in the followingdocuments, which are incorporated by reference in their entirety: U.S.Application Publication No. 2010/0076057 (Sontheimer et al., Target DNAInterference with crRNA); U.S. Application Publication No. 2014/0179006(Feng, CRISPR-CAS Component Systems, Methods, and Compositions forSequence Manipulation); U.S. Application Publication No. 2014/0294773(Brouns et al., Modified Cascade Ribonucleoproteins and Uses Thereof);Sorek et al., Annu. Rev. Biochem. 82:273-266, 2013; and Wang, S. et al.,Plant Cell Rep (2015) 34: 1473-1476. Therefore, it is another embodimentto use the CRISPR system on hybrid pepper XHP16823 to modify traits andtolerances to pests, herbicides, and viruses.

Gene Editing Using TALENs

Transcription activator-like effector nucleases (TALENs) have beensuccessfully used to introduce targeted mutations via repair of doublestranded breaks (DSBs) either through non-homologous end joining (NHEJ),or by homology-directed repair (HDR) and homology-independent repair inthe presence of a donor template. Thus, TALENs are another mechanism fortargeted genome editing using XHP16823. The technique is well known inthe art; see for example Malzahn, Aimee et al. “Plant genome editingwith TALEN and CRISPR” Cell & bioscience vol. 7 21. 24 Apr. 2017.

Therefore, it is another embodiment to use the TALENs system on hybridpepper XHP16823 to modify traits and resistances or tolerances to pests,herbicides, and viruses.

Other Methods of Genome Editing

In addition to CRISPR and TALENs, two other types of engineerednucleases can be used for genome editing: engineered homingendonucleases/meganucleases (EMNs), and zinc finger nucleases (ZFNs).These methods are well known in the art. See for example, Petilino,Joseph F. “Genome editing in plants via designed zinc finger nucleases”In Vitro Cell Dev Biol Plant. 51(1): pp. 1-8 (2015); and Daboussi,Fayza, et al. “Engineering Meganuclease for Precise Plant GenomeModification” in Advances in New Technology for Targeted Modification ofPlant Genomes. Springer Science+Business. pp 21-38 (2015).

Therefore, it is another embodiment to use engineered nucleases onhybrid pepper XHP16823 to modify traits and resistances or tolerances topests, herbicides, and viruses.

Introduction of a New Trait or Locus Into Hybrid Pepper XHP16823

Hybrid pepper XHP16823 represents a new variety into which a new locusor trait may be introgressed. Direct transformation and backcrossingrepresent two important methods that can be used to accomplish such anintrogression. The term backcross conversion and single locus conversionare used interchangeably to designate the product of a backcrossingprogram.

Molecular Techniques Using Hybrid Pepper XHP16823

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions, such as encoding specific protein products. Scientists in thefield of plant biology developed a strong interest in engineering thegenome of plants to contain and express foreign genetic elements, oradditional, or modified versions of native or endogenous geneticelements in order to “alter” (the utilization of up-regulation,down-regulation, or gene silencing) the traits of a plant in a specificmanner. Any DNA sequences, whether from a different species or from thesame species, which are introduced into the genome using transformationor various breeding methods are referred to herein collectively as“transgenes” and are known as one gene editing technique. In someembodiments, a transgenic variant of hybrid pepper XHP16823 may containat least one transgene. Over the last fifteen to twenty years severalmethods for producing transgenic plants have been developed, and anotherembodiment also relates to transgenic variants of the claimed hybridpepper XHP16823.

Nucleic acids or polynucleotides refer to RNA or DNA that is linear orbranched, single or double stranded, or a hybrid thereof. The term alsoencompasses RNA/DNA hybrids. These terms also encompass untranslatedsequence located at both the 3′ and 5′ ends of the coding region of thegene: at least about 1000 nucleotides of sequence upstream from the 5′end of the coding region and at least about 200 nucleotides of sequencedownstream from the 3′ end of the coding region of the gene. Less commonbases, such as inosine, 5-methylcytosine, 6-methyladenine, hypoxanthineand others can also be used for antisense, dsRNA and ribozyme pairing.For example, polynucleotides that contain C-5 propyne analogues ofuridine and cytidine have been shown to bind RNA with high affinity andto be potent antisense inhibitors of gene expression. Othermodifications, such as modification to the phosphodiester backbone, orthe 2′-hydroxy in the ribose sugar group of the RNA can also be made.The antisense polynucleotides and ribozymes can consist entirely ofribonucleotides, or can contain mixed ribonucleotides anddeoxyribonucleotides. The polynucleotides of the embodiments may beproduced by any means, including genomic preparations, cDNApreparations, in-vitro synthesis, RT-PCR, and in vitro or in vivotranscription.

One embodiment is a process for producing hybrid pepper XHP16823 furthercomprising a desired trait, said process comprising introducing atransgene that confers a desired trait to a pepper plant of hybridpepper XHP16823. Another embodiment is the product produced by thisprocess. In one embodiment, the desired trait may be one or more ofherbicide tolerance, insect tolerance, disease tolerance, environmentalstress tolerance, or modified carbohydrate metabolism. The specific genemay be any known in the art or listed herein, including: apolynucleotide conferring resistance to imidazolinone, dicamba,sulfonylurea, glyphosate, glufosinate, triazine, PPO-inhibitorherbicides, benzonitrile, cyclohexanedione, phenoxy proprionic acid, andL-phosphinothricin; a polynucleotide encoding a Bacillus thuringiensispolypeptide; or a polynucleotide conferring resistance to Tomato spottedwilt virus, Xanthomonas euvesicatoria, Bs2, CARAV1, or CaPMEI1.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Li D. et al, “Establishment of a highly efficienttransformation system for pepper (Capsicum annuum L.)” Plant CellReports, pp 785-788 (2003), and Lee Y. H. et al, “A new selection methodfor pepper transformation: callus-mediated shoot formation” Plant CellReports pp 50-58 (2004). In addition, expression vectors and in vitroculture methods for plant cell or tissue transformation and regenerationof plants are available. See, for example, Gruber, et al., “Vectors forPlant Transformation,” in Methods in Plant Molecular Biology andBiotechnology, Glick and Thompson Eds., CRC Press, Inc., Boca Raton, pp.89-119 (1993) and Nakagawa T. et al, “Development of series of gatewaybinary vectors, pGWBs, for realizing efficient construction of fusiongenes for plant transformation” Journal of Bioscience and Bioengineeringpp 34-41 (2007).

A genetic trait which has been engineered into the genome of aparticular pepper plant may then be moved into the genome of anothervariety using traditional breeding techniques that are well known in theplant breeding arts. For example, a backcrossing approach is commonlyused to move a transgene from a transformed hybrid pepper line into analready developed hybrid pepper line, and the resulting backcrossconversion plant would then comprise the transgene(s).

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include, but are not limited to, genes,coding sequences, inducible, constitutive and tissue specific promoters,enhancing sequences, and signal and targeting sequences. For example,see the traits, genes, and transformation methods listed in U.S. Pat.No. 6,118,055.

Breeding with Molecular Markers

Molecular markers, which includes markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Amplified Fragment Length Polymorphisms (AFLPs), Arbitrarily PrimedPolymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs), Simple SequenceRepeats (SSRs), and Single Nucleotide Polymorphisms (SNPs) may be usedin plant breeding methods utilizing hybrid pepper XHP16823.

Isozyme Electrophoresis and RFLPs have been widely used to determinegenetic composition. See Lee J. M et al, “Characterization and moleculargenetic mapping of microsatellite loci in pepper” Theoretical andApplied Genetics pp 619-627 (2004), Kang B. C. et al, “An interspecific(Capsicum annuum x C. Chinese) F2 linkage map in pepper using RFLP andALFP markers” Theoretical and Applied Genetics pp 531-539 (2001), andLefebvre V. et al, “Nuclear RFLP between pepper cultivars (Capsicumannuum L.) Euphytica pp 189-199 (1993).

Randomly Amplified Polymorphic DNAs (RAPDs) and Amplified FragmentLength Polymorphisms (AFLPs) may also be used. See for example Paran, I.et al, “Variation in Capsicum annuum revealed by RAPD and AFLP markers”Euphytica, pp 167-173 (1998), and Llbi, H. “RAPD markers assistedvarietal identification and genetic purity test in pepper, Capsicumannuum” Scientia Horticulturae, pp 211-218 (2003).

SSR technology can be routinely used. See Kwon, Y. et al, “Use of SSRMarkers to Complement Tests of Distinctiveness, Uniformity, andStability (DUS) of Pepper (Capsicum annuum L.) Varieties, Mol. Cells, pp1-8 (2005) and Minamiyama, Y., et al “An SSR-based linkage map ofCapsicum annuum” Molecular Breeding, pp 157-169 (2006).

Single Nucleotide Polymorphisms (SNPs) may also be used to identify theunique genetic composition of the embodiment(s) and progeny varietiesretaining that unique genetic composition. See Jung, J. et al “Discoveryof single nucleotide polymorphism in Capsicum and SNP markers forcultivar identification” Euphytica pp 91-107 (2010), Nicolai, M., et al“Discovery of a large set of SNP and SSR genetic markers byhigh-throughput sequencing of pepper (Capsicum annuum)” Genetics andMolecular Research, pp 2295-2300 (2012), Jeong, H. et al,“Identification of Capsicum species using SNP markers based on highresolution melting analysis” Genome pp 1029-1040 (2010), and Ashrafi,H., et al, “De novo assembly of the pepper transcriptome (Capsicumannuum): a benchmark for in silico discovery of SNPs, SSRs and candidategenes” BMC Genomics (2012).

One use of molecular markers is Quantitative Trait Loci (QTL) mapping.QTL mapping is the use of markers, which are known to be closely linkedto alleles that have measurable effects on a quantitative trait.Selection in the breeding process is based upon the accumulation ofmarkers linked to the positive effecting alleles and/or the eliminationof the markers linked to the negative effecting alleles from the plant'sgenome. See Chaim, A. B., et al, “QTL mapping of fruit-related traits inpepper (Capsicum annuum), Theoretical and Applied Genetics, pp 1016-1028(2001) and Rao, G. U., et al, “Mapping of yield-related QTLs in pepperin an interspecific cross of Capsicum annuum and C. frutescens”Theoretical and Applied Genetics, pp 1457-1466 (2003). QTL markers canalso be used during the breeding process for the selection ofqualitative traits. For example, markers closely linked to alleles ormarkers containing sequences within the actual alleles of interest canbe used to select plants that contain the alleles of interest during abackcrossing breeding program. The markers can also be used to selectfor the genome of the recurrent parent and against the genome of thedonor parent. Using this procedure can minimize the amount of genomefrom the donor parent that remains in the selected plants. It can alsobe used to reduce the number of crosses back to the recurrent parentneeded in a backcrossing program. The use of molecular markers in theselection process is often called genetic marker enhanced selection.Molecular markers may also be used to identify and exclude certainsources of germplasm as parental varieties or ancestors of a plant byproviding a means of tracking genetic profiles through crosses.

Production of Double Haploids

The production of double haploids can also be used for the developmentof plants with a homozygous phenotype in the breeding program. Forexample, a pepper plant for which hybrid pepper XHP16823 is a parent canbe used to produce double haploid plants. Double haploids are producedby the doubling of a set of chromosomes (1N) from a heterozygous plantto produce a completely homozygous individual. This can be advantageousbecause the process omits the generations of selfing needed to obtain ahomozygous plant from a heterozygous source. For example, see, Keleş, D.et al, “Effect of Pepper Types on Obtaining Spontaneous Doubled HaploidPlants via Anther culture” HortScience, pp1671-1676 (2015), Olszewska,D. et al, “The assessment of doubled haploid lines obtained in pepper(Capsicum annuum L.) anther culture” Folia Horticulturae, pp 93-99(2011), and M. Maluszynski et al. (eds), Doubled Haploid Production inCrop Plants, (2003).

Thus, an embodiment is a process for making a substantially homozygoushybrid pepper XHP16823 progeny plant by producing or obtaining a seedfrom the cross of hybrid pepper XHP16823 and another pepper plant andapplying double haploid methods to the F₁ seed or F₁ plant or to anysuccessive filial generation.

In particular, a process of making seed retaining the molecular markerprofile of hybrid pepper XHP16823 is contemplated, such processcomprising obtaining or producing F₁ seed for which hybrid pepperXHP16823 is a parent, inducing doubled haploids to create progenywithout the occurrence of meiotic segregation, obtaining the molecularmarker profile of hybrid pepper XHP16823, and selecting progeny thatretain the molecular marker profile of hybrid pepper XHP16823.

Expression Vectors for Pepper Transformation: Marker Genes

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to, aregulatory element (for example, a promoter). Expression vectors includeat least one genetic marker operably linked to a regulatory element (forexample, a promoter) that allows transformed cells containing the markerto be either recovered by negative selection, i.e., inhibiting growth ofcells that do not contain the selectable marker gene, or by positiveselection, i.e., screening for the product encoded by the geneticmarker. Many commonly used selectable marker genes for planttransformation are well-known in the transformation arts, and include,for example, genes that code for enzymes that metabolically detoxify aselective chemical agent which may be an antibiotic or an herbicide, orgenes that encode an altered target which is insensitive to theinhibitor. A few positive selection methods are also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene which, when under thecontrol of plant regulatory signals, confers resistance to kanamycin.Another commonly used selectable marker gene is the hygromycinphosphotransferase gene which confers resistance to the antibiotichygromycin.

Additional selectable marker genes include Pain1-9a and Pain1-8c whichboth correspond to the group a alleles of the vacuolar acid invertasegene; Pain1prom-d/e; Stp23-8b, StpL-3b, and StpL-3e which originate fromtwo plastid starch phosphorylase genes; AGPsS-9a which is positivelyassociated an increase in plant starch content, starch yield and chipquality, and AGPsS-10a which is associated with a decrease in theaverage plant starch content, starch yield and chip quality; GP171-awhich corresponds to allele 1a of ribulose bisphosphate carboxylaseactivase; and Rca-1a.

Selectable marker genes for plant transformation not of bacterial origininclude, for example, mouse dihydrofolate reductase, plant5-enolpyruvylshikimate-3-phosphate synthase, and plant acetolactatesynthase (Eichholtz, et al., Somatic Cell Mol. Genet., 13:67 (1987);Shah, et al., Science, 233:478 (1986); Charest, et al., Plant Cell Rep.,8:643 (1990)).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells, rather than directgenetic selection of transformed cells, for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used marker genes forscreening presumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase, and chloramphenicol acetyltransferase(Jefferson, R. A., Plant Mol. Biol. Rep., 5:387 (1987); Teeri, et al.,EMBO J., 8:343 (1989); Koncz, et al., Proc. Natl. Acad. Sci. USA, 84:131(1987); DeBlock, et al., EMBO J., 3:1681 (1984)).

Expression Vectors for Pepper Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element (for example, a promoter).Several types of promoters are well known in the transformation arts asare other regulatory elements that can be used alone or in combinationwith promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters that initiate transcription only in a certain tissue arereferred to as “tissue-specific.” A “cell-type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may affect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell-type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter that is active under mostenvironmental conditions. Many types of promoters are well known in theart.

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of a protein produced by transgenes to a subcellularcompartment, such as the chloroplast, vacuole, peroxisome, glyoxysome,cell wall, or mitochondrion, or for secretion into the apoplast, isaccomplished by means of operably linking the nucleotide sequenceencoding a signal sequence to the 5′ and/or 3′ region of a gene encodingthe protein of interest. Targeting sequences at the 5′ and/or 3′ end ofthe structural gene may determine during protein synthesis andprocessing where the encoded protein is ultimately compartmentalized.Many signal sequences are well-known in the art. See, for example,Becker, et al., Plant Mol. Biol., 20:49 (1992); Knox, C., et al., PlantMol. Biol., 9:3-17 (1987); Lerner, et al., Plant Physiol., 91:124-129(1989); Frontes, et al., Plant Cell, 3:483-496 (1991); Matsuoka, et al.,Proc. Natl. Acad. Sci., 88:834 (1991); Gould, et al., J. Cell. Biol.,108:1657 (1989); Creissen, et al., Plant J., 2:129 (1991); Kalderon, etal., Cell, 39:499-509 (1984); Steifel, et al., Plant Cell, 2:785-793(1990).

Foreign Protein Genes and Agronomic Genes: Transformation

With transgenic plants, according to one embodiment, a foreign proteincan be produced in commercial quantities. Thus, techniques for theselection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein can then beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem., 114:92-6(1981).

According to an embodiment, the transgenic plant provided for commercialproduction of foreign protein is a pepper plant. In another embodiment,the biomass of interest is seed. For the relatively small number oftransgenic plants that show higher levels of expression, a genetic mapcan be generated, primarily via conventional RFLP, PCR, and SSRanalysis, which identifies the approximate chromosomal location of theintegrated DNA molecule. For exemplary methodologies in this regard,see, Glick and Thompson, Methods in Plant Molecular Biology andBiotechnology, CRC Press, Inc., Boca Raton, 269:284 (1993). Mapinformation concerning chromosomal location is useful for proprietaryprotection of a subject transgenic plant.

Likewise, by means of one embodiment, plants can be geneticallyengineered to express various phenotypes of agronomic interest. Throughthe transformation of pepper, the expression of genes can be altered toenhance disease tolerance, insect tolerance, herbicide tolerance,agronomic, grain quality, and other traits. Transformation can also beused to insert DNA sequences which control or help controlmale-sterility. DNA sequences native to peppers, as well as non-nativeDNA sequences, can be transformed into peppers and used to alter levelsof native or non-native proteins. Various promoters, targetingsequences, enhancing sequences, and other DNA sequences can be insertedinto the genome for the purpose of altering the expression of proteins.The interruption or suppression of the expression of a gene at the levelof transcription or translation (also known as gene silencing or genesuppression) is desirable for several aspects of genetic engineering inplants.

Many techniques for gene silencing are well-known to one of skill in theart, including, but not limited to, knock-outs (such as by insertion ofa transposable element such as Mu (Vicki Chandler, The Maize Handbook,Ch. 118 (Springer-Verlag 1994)) or other genetic elements such as a FRT,Lox, or other site specific integration sites; antisense technology(see, e.g., Sheehy, et al., PNAS USA, 85:8805-8809 (1988) and U.S. Pat.Nos. 5,107,065, 5,453,566, and 5,759,829); co-suppression (e.g., Taylor,Plant Cell, 9:1245 (1997); Jorgensen, Trends Biotech., 8(12):340-344(1990); Flavell, PNAS USA, 91:3490-3496 (1994); Finnegan, et al.,Bio/Technology, 12:883-888 (1994); Neuhuber, et al., Mol. Gen. Genet.,244:230-241 (1994)); RNA interference (Napoli, et al., Plant Cell,2:279-289 (1990); U.S. Pat. No. 5,034,323; Sharp, Genes Dev., 13:139-141(1999); Zamore, et al., Cell, 101:25-33 (2000); Montgomery, et al., PNASUSA, 95:15502-15507 (1998)), virus-induced gene silencing (Burton, etal., Plant Cell, 12:691-705 (2000); Baulcombe, Curr. Op. Plant Bio.,2:109-113 (1999)); target-RNA-specific ribozymes (Haseloff, et al.,Nature, 334:585-591 (1988)); hairpin structures (Smith, et al., Nature,407:319-320 (2000); U.S. Pat. Nos. 6,423,885, 7,138,565, 6,753,139, and7,713,715); MicroRNA (Aukerman & Sakai, Plant Cell, 15:2730-2741(2003)); ribozymes (Steinecke, et al., EMBO J., 11:1525 (1992);Perriman, et al., Antisense Res. Dev., 3:253 (1993)); oligonucleotidemediated targeted modification (e.g., U.S. Pat. Nos. 6,528,700 and6,911,575); Zn-finger targeted molecules (e.g., U.S. Pat. Nos.7,151,201, 6,453,242, 6,785,613, 7,177,766 and 7,788,044); and othermethods or combinations of the above methods known to those of skill inthe art.

The foregoing methods for transformation may be used for producing atransgenic variety. The transgenic variety could then be crossed withanother (non-transformed or transformed) variety in order to produce anew transgenic variety. Alternatively, a genetic trait that has beenengineered into a particular pepper line using the foregoingtransformation techniques could be moved into another line usingtraditional backcrossing techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove an engineered trait from a public, non-elite variety into an elitevariety, or from a variety containing a foreign gene in its genome intoa variety or varieties that do not contain that gene. As used herein,“crossing” can refer to a simple x by y cross or the process ofbackcrossing depending on the context.

Likewise, by means of one embodiment, agronomic genes can be expressedin transformed plants. More particularly, plants can be geneticallyengineered to express various phenotypes of agronomic interest,including, but not limited to, genes that confer resistance to pests ordisease, genes that confer resistance to an herbicide, genes that conferor contribute to a value-added or desired trait, genes that control malesterility, genes that create a site for site specific DNA integration,and genes that affect abiotic stress resistance. Many hundreds if notthousands of different genes are known and could potentially beintroduced into a pepper plant according to the invention. Non-limitingexamples of particular genes and corresponding phenotypes one may chooseto introduce into a pepper plant include one or more genes for insecttolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerancesuch as genes for fungal disease control, herbicide tolerance such asgenes conferring glyphosate tolerance, and genes for qualityimprovements such as yield, nutritional enhancements, environmental orstress tolerances, or any desirable changes in plant physiology, growth,development, morphology or plant product(s). For example, structuralgenes would include any gene that confers insect tolerance including butnot limited to a Bacillus insect control protein gene as described in WO99/31248, herein incorporated by reference in its entirety, U.S. Pat.No. 5,689,052, herein incorporated by reference in its entirety, U.S.Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference intheir entirety. In another embodiment, the structural gene can confertolerance to the herbicide glyphosate as conferred by genes including,but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPSgene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, hereinincorporated by reference in its entirety, or glyphosate oxidoreductasegene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporatedby reference in its entirety. Alternatively, the DNA coding sequencescan affect these phenotypes by encoding a non-translatable RNA moleculethat causes the targeted inhibition of expression of an endogenous gene,for example via antisense- or cosuppression-mediated mechanisms (see,for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991). TheRNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineeredto cleave a desired endogenous mRNA product (see for example, Gibson andShillito, Mol. Biotech., 7:125, 1997). Thus, any gene which produces aprotein or mRNA which expresses a phenotype or morphology change ofinterest is useful for the practice of one or more embodiments.

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of peppers andregeneration of plants therefrom is well-known and widely published.See, Agrawal, S. et al., “Plant regeneration in tissue cultures ofpepper (Capsicum annuum L. cv. Mathania) Plant Cell, Tissue and OrganCulture, Vol. 16(1) pp 47-55 1989; Berljak J. “In vitro plantregeneration from Pepper (Capsicum annuum L. cv. ‘Soroksari’) SeedlingExplants” Phyton (Austria) Special issue: “Plant Physiology” Vol. 39(3)pp 289-292 (1999); Ahmad, N., et al., “Improved plant regeneration inCapsicum annuum L. from nodal segments” Biologia Plantarum Vol. 50(4) pp701-704 (2006); and Otroshy, M. et al., “Micropropagation of Pepper(Capsicum annuum L.) Through in vitro Direct Organogenesis” AsianJournal of Biotechnology Vol. 3 pp 38-45 (2010). Thus, another aspect orembodiment is to provide cells which upon growth and differentiationproduce pepper plants having the physiological and morphologicalcharacteristics of hybrid pepper XHP16823.

Regeneration refers to the development of a plant from tissue culture.The term “tissue culture” indicates a composition comprising isolatedcells of the same or a different type or a collection of such cellsorganized into parts of a plant. Exemplary types of tissue cultures areprotoplasts, calli, plant clumps, and plant cells that can generatetissue culture that are intact in plants or parts of plants, such asembryos, pollen, flowers, seeds, pods, petioles, leaves, stems, roots,root tips, anthers, pistils, and the like. Means for preparing andmaintaining plant tissue culture are well known in the art. By way ofexample, a tissue culture comprising organs has been used to produceregenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and 5,977,445describe certain techniques, the disclosures of which are incorporatedherein by reference.

Industrial Uses

Pepper has a wide variety of uses in the commodity area. For example,fresh peppers can be eaten raw or cooked (fried, baked, boiled, etc.).Peppers can also be used to make spices. Thus, a further embodimentprovides for a food product made from a part of the pepper plant varietyXHP16823.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

One embodiment may be embodied in other specific forms without departingfrom its spirit or essential characteristics. The described embodimentsare to be considered in all respects only as illustrative and notrestrictive. All changes which come within the meaning and range ofequivalency of the claims are to be embraced within their scope.

Various embodiments, include components, methods, processes, systemsand/or apparatus substantially as depicted and described herein,including various embodiments, sub-combinations, and subsets thereof.Those of skill in the art will understand how to make and use anembodiment(s) after understanding the present disclosure.

The foregoing discussion of the embodiments has been presented forpurposes of illustration and description. The foregoing is not intendedto limit the embodiments to the form or forms disclosed herein. In theforegoing Detailed Description for example, various features of theembodiments are grouped together in one or more embodiments for thepurpose of streamlining the disclosure. This method of disclosure is notto be interpreted as reflecting an intention that the embodiment(s)requires more features than are expressly recited in each claim. Rather,as the following claims reflect, inventive aspects lie in less than allfeatures of a single foregoing disclosed embodiment. Thus, the followingclaims are hereby incorporated into the detailed description.

Moreover, though the description of the embodiments has includeddescription of one or more embodiments and certain variations andmodifications, other variations and modifications are within the scopeof the embodiments (e.g., as may be within the skill and knowledge ofthose in the art, after understanding the present disclosure). It isintended to obtain rights which include alternative embodiments to theextent permitted, including alternate, interchangeable and/or equivalentstructures, functions, ranges or acts to those claimed, whether or notsuch alternate, interchangeable and/or equivalent structures, functions,ranges or acts are disclosed herein, and without intending to publiclydedicate any patentable subject matter.

The use of the terms “a,” “an,” and “the,” and similar referents in thecontext of describing the embodiments (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the embodiments and does not pose a limitation on the scopeof the embodiments unless otherwise claimed.

Deposit Information

A deposit of proprietary hybrid pepper seed XHP16823 disclosed above andrecited in the appended claims is being maintained by Sakata SeedAmerica, Inc. A deposit will be made with the Provasoli-GuillardNational Center for Marine Algae and Microbiota (NCMA), BigelowLaboratory for Ocean Sciences, 60 Bigelow Drive, East Boothbay, Me.04544, United States. Access to this deposit will be available duringthe pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to a deposit ofthe seed deposit of the same variety with NCMA. The deposit will bemaintained in the depository for a period of 30 years, or 5 years afterthe last request, or for the effective life of the patent, whichever islonger, and will be replaced if necessary, during that period.

1. A seed of hybrid pepper plant designated XHP16823, wherein a sampleof seed of hybrid pepper XHP16823 was deposited under NCMA No.202103006.
 2. A pepper plant produced from the seed of claim
 1. 3. Aplant part of the plant of claim 2, wherein said part is a cell, pollen,leaf, ovule, anther, cotyledon, hypocotyl, pistil, root, root tip,flower, fruit, petiole, or stem.
 4. A tissue culture produced fromprotoplasts or cells from the plant of claim 2, wherein said cells orprotoplasts are produced from a plant part selected from the groupconsisting of embryo, leaf, pollen, ovule, cotyledon, hypocotyl,meristematic cell, callus, root, root tip, pistil, anther, flower,fruit, shoot, stem, and petiole.
 5. A pepper plant produced from thetissue culture of claim 4, having all of the physiological andmorphological characteristics of hybrid pepper XHP16823.
 6. A method ofvegetatively propagating the pepper plant of claim 2, comprising thesteps of: (a) collecting tissue capable of being propagated from theplant according to claim 2; (b) cultivating said tissue to obtainproliferated shoots; and (c) rooting said proliferated shoots to obtainrooted plantlets.
 7. The method of claim 6, further comprising growingat least a first pepper plant from said rooted plantlets.
 8. A methodfor producing a seed of a pepper plant derived from pepper hybridXHP16823, comprising the steps of: (a) selfing a pepper plant of hybridXHP16823 with itself or crossing a pepper plant of hybrid XHP16823 witha second pepper plant, wherein a sample of seed of hybrid XHP16823 wasdeposited under NCMA No. 202103006; and (b) allowing seed of a hybridXHP16823-derived pepper plant to form.
 9. The method of claim 13,further comprising the steps of: (c) selfing a plant grown from saidhybrid XHP16823-derived pepper seed to yield additional hybridXHP16823-derived pepper seed; (d) growing said additional hybridXHP16823-derived pepper seed of step (c) to yield additional hybridXHP16823-derived pepper plants; and (e) repeating the selfing andgrowing steps of (c) and (d) to generate at least a first further hybridXHP16823-derived pepper plant.
 10. The method of claim 14, furthercomprising: (f) crossing the further hybrid XHP16823-derived pepperplant with a second pepper plant to produce seed of a hybrid progenyplant.
 11. A method of producing a pepper fruit, comprising: (a)obtaining the plant according to claim 2, wherein the plant has beencultivated to maturity; and (b) collecting a pepper fruit from saidplant.
 12. The method of claim 11, wherein said second pepper plant isan inbred pepper line.
 13. A plant part of the plant of claim 2, whereinthe plant part comprises a cell of said hybrid pepper XHP16823.
 14. Amethod for developing a pepper plant in a plant breeding program,comprising applying plant breeding techniques to the plant, or plantpart thereof, of claim 2, comprising crossing, recurrent selection,mutation breeding, wherein said mutation breeding selects for a mutationthat is spontaneous or artificially induced, backcrossing, pedigreebreeding, marker enhanced selection, haploid/double haploid production,or transformation, wherein application of said techniques results indevelopment of a pepper plant.
 15. A method of introducing a mutationinto the genome of pepper plant XHP16823, said method comprisingmutagenesis of the plant, or plant part thereof, of claim 2, whereinsaid mutagenesis is selected from the group consisting of temperature,long-term seed storage, tissue culture conditions, ionizing radiation,chemical mutagens, or targeting induced local lesions in genomes, andwherein the resulting plant comprises at least one genome mutation. 16.A method of editing the genome of pepper plant XHP16823, said methodcomprising editing the genome of the plant, or plant part thereof,wherein a sample of seed of hybrid XHP16823 plant was deposited underNCMA No. 202103006, wherein said method is selected from the groupcomprising transgene, zinc finger nucleases, transcriptionactivator-like effector nucleases (TALENs), engineered homingendonucleases/meganucleases, and the clustered regularly interspacedshort palindromic repeat (CRISPR)-associated protein9 (Cas9) system. 17.A pepper plant produced by the method of claim 16, wherein said planthas all of the physiological and morphological characteristics of hybridpepper XHP16823.
 18. The method of claim 16, wherein the gene editingresults in conferring a desired trait, wherein the desired traitselected from the group consisting of male sterility, herbicidetolerance, insect tolerance, pest tolerance, disease tolerance, modifiedfatty acid metabolism, environmental stress tolerance, and modifiedcarbohydrate metabolism.
 19. The method of claim 18, wherein the traitis herbicide tolerance and wherein said herbicide tolerance is conferredto an herbicide selected from the group consisting of imidazolinone,sulfonylurea, glyphosate, glufosinate, L-phosphinothricin, triazine andbenzonitrile.
 20. A pepper plant produced by the method of claim 18,wherein said plant has all of the physiological and morphologicalcharacteristics of hybrid pepper XHP16823.
 21. A pepper plant producedby the method of claim 19, wherein said plant has all of thephysiological and morphological characteristics of hybrid pepperXHP16823.